TCF1/TCF7 Antibody Performs Well in Sub-cellular Fractionation

September 29, 2026

BlueskyReddit
Biochemistry
Stanford University
Senior Research Scientist

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Company:

Cell Signaling

Product Name:

TCF1/TCF7 (C63D9) Rabbit Monoclonal Antibody

Catalog Number:

2203

Clone Number:

C63D9

WNT signaling is essential for embryonic development and adult tissue homeostasis, and its aberrant activation drives several cancers, most notably colorectal cancer. My lab studies the molecular mechanisms of oncogenic WNT signal transduction, with the goal of identifying therapeutic vulnerabilities in colorectal cancer. Stabilized β-catenin acts in the nucleus with TCF/LEF transcription factors, so we use subcellular fractionation to follow pathway components across compartments during WNT stimulation.

Experimental Design and Results Summary

Applications

Western Blot

Sample

HEK293T cytoplasmic and nuclear fractions

Primary Incubation

1:1000

Blocking Agent

5% milk in TBS-T (0.1% Tween-20)

Secondary Incubation

HRP-conjugated Donkey anti-Rabbit IgG; 1:10,000 dilution

Tertiary Incubation

None

Detection

ECL picoplus/ECL femto

Results Summary

We treated HEK293T cells with 50% WNT3A-conditioned medium for 0, 0.5, 1, 2, 4, and 8 h, then separated cytosolic and nuclear fractions by hypotonic swelling, needle shearing, and differential centrifugation. Cytosolic fractions (30 µg) and equal volumes of the corresponding nuclear fractions, representing approximately equal cell equivalents, were resolved on 4–12% Bis-Tris gels and transferred to nitrocellulose. Membranes were blocked in 5% milk/TBS-T, probed with the TCF1/TCF7 antibody at 1:1000 overnight at 4 °C, then with HRP-conjugated donkey anti-rabbit IgG (1:10,000) and ECL picoplus/ECL femto. The antibody detected a single sharp band at approximately 45 kDa, present only in nuclear fractions and undetectable in all cytoplasmic lanes. α-Tubulin blotted from the same fractions was restricted to the cytoplasm, supporting the conclusion that the nuclear signal reflects genuine localization. We did not include a TCF1/TCF7 knockout or knockdown control, so band identity was not confirmed genetically.

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Summary

The Good

The antibody gives a very clean result: one sharp band, no background across the blot, and no signal in cytoplasmic fractions. Endogenous TCF1 was readily detectable in 293T cells at 1:1000 in milk-based buffer. The nuclear-restricted pattern is consistent with TCF1's role as a DNA-binding transcription factor and makes the antibody well suited to fractionation and nuclear-translocation studies.

The Bad

None

The Bottom Line

Cell Signaling's TCF1/TCF7 antibody is a specific-looking reagent for detecting endogenous TCF1/TCF7 by western blot, and it performed very well in sub-cellular fractionation. I recommend it, with the caveat that specificity was not confirmed with a TCF1/TCF7-null control.

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